WO2023022269A1 - Pharmaceutical composition for preventing or treating attention deficit/hyperactivity disorder - Google Patents

Pharmaceutical composition for preventing or treating attention deficit/hyperactivity disorder Download PDF

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WO2023022269A1
WO2023022269A1 PCT/KR2021/011136 KR2021011136W WO2023022269A1 WO 2023022269 A1 WO2023022269 A1 WO 2023022269A1 KR 2021011136 W KR2021011136 W KR 2021011136W WO 2023022269 A1 WO2023022269 A1 WO 2023022269A1
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Prior art keywords
adhd
attention deficit
hyperactivity disorder
pharmaceutical composition
deficit hyperactivity
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PCT/KR2021/011136
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French (fr)
Korean (ko)
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윤보은
이창준
김유성
사문선
Original Assignee
단국대학교 천안캠퍼스 산학협력단
기초과학연구원
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Application filed by 단국대학교 천안캠퍼스 산학협력단, 기초과학연구원 filed Critical 단국대학교 천안캠퍼스 산학협력단
Priority to EP21954316.2A priority Critical patent/EP4389126A1/en
Priority claimed from KR1020210110120A external-priority patent/KR102597711B1/en
Priority claimed from KR1020210110121A external-priority patent/KR102684933B1/en
Publication of WO2023022269A1 publication Critical patent/WO2023022269A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids

Definitions

  • the present invention relates to a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD).
  • ADHD attention deficit hyperactivity disorder
  • ADHD Attention deficit hyperactivity disorder
  • appetite suppressants such as amphetamines, methylphenidate, and pemoline.
  • antidepressants such as desipramine, which act to selectively block the reuptake of norepinephrine, are also effective in some cases.
  • Newer drugs such as atomoxetine that block the reuptake of norepinephrine and serotonin may also be effective in treating this disorder.
  • Psychostimulants and monoamine reuptake inhibitors control activity levels and attention, but are ineffective in treating the comorbid or concomitant cognitive deficits associated with ADHD.
  • methylphenidate (MPH)
  • ADHD symptoms are related to the control of noradrenergic neurons that control neural circuits in the frontal lobe.
  • noradrenergic and dopaminergic neurons Studies have also reported that the behavior of ADHD is caused by an imbalance between the control of noradrenergic and dopaminergic neurons.
  • catecholamines which control glutamatergic neurons and GABA (g-aminobutyric acid) neurons, on the post-synaptic area is reduced.
  • An object of the present invention is to provide a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) containing a component exhibiting an effect capable of improving hyperactivity disorder.
  • ADHD attention deficit hyperactivity disorder
  • Another object of the present invention is to provide a biomarker composition for diagnosing attention deficit hyperactivity disorder (ADHD).
  • ADHD attention deficit hyperactivity disorder
  • the present invention provides a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) comprising SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient.
  • ADHD attention deficit hyperactivity disorder
  • the present invention provides a preventive or functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and food additives acceptable food additives.
  • ADHD attention deficit hyperactivity disorder
  • the present invention provides a biomarker composition for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising GAT-3 (GABA transporter-3) or a gene encoding the same as an active ingredient.
  • ADHD attention deficit hyperactivity disorder
  • the present invention provides a diagnostic kit for attention deficit hyperactivity disorder (ADHD) comprising, as an active ingredient, an agent capable of detecting the expression level of GAT-3 (GABA transporter-3) protein or the expression level of a gene encoding the same. to provide.
  • ADHD attention deficit hyperactivity disorder
  • the present invention measures the expression level of GAT-3 (GABA transporter-3) protein or the expression level of the gene encoding it in a biological sample isolated from a subject suspected of having attention deficit hyperactivity disorder (ADHD); And it provides an information providing method for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising the step of comparing the expression level with a biological sample isolated from a normal person.
  • GAT-3 GABA transporter-3
  • ADHD attention deficit hyperactivity disorder
  • the present invention it was confirmed that the amount of GABA increased in the striatum of the brain, which is in charge of hyperactivity, in Git1 gene-defective hetero (+/-) mice, thereby preventing attention deficit/hyperactivity disorder.
  • disorder, ADHD can be used as an animal model, and by confirming that hyperactivity is improved by administering SNAP5114 to Git1 gene-defective hetero (+/-) mice, SNAP5114 can be used as an attention deficit/hyperactivity disorder (ADHD) It can be provided as a treatment for hyperactivity disorder (ADHD), and GABA transporter-3 (GAT-3) can be provided as a biomarker for diagnosing attention deficit/hyperactivity disorder (ADHD).
  • 3 is a result of evaluating hyperactivity changes according to SNAP5114 administration in an ADHD (attention deficit / hyperactivity disorder) animal model.
  • the present invention provides a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) comprising SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient.
  • ADHD attention deficit hyperactivity disorder
  • the SNAP5114 is a compound represented by Formula 1 below, specifically (S)-1-(2-(tris(4-methoxyphenyl)methoxy)ethyl)piperidine-3-carboxylic acid ((S) -1-(2-(tris(4-methoxyphenyl)methoxy)ethyl)piperidine-3-carboxylic acid).
  • the SNAP5114 has an effect of improving hyperactivity in an attention deficit / hyperactivity disorder (ADHD) animal model.
  • ADHD attention deficit / hyperactivity disorder
  • the pharmaceutically acceptable salt refers to an acid addition salt formed by a pharmaceutically acceptable free acid
  • the pharmaceutically acceptable salt refers to a salt commonly used in the pharmaceutical industry, for example
  • an inorganic ion salt made of calcium, potassium, sodium or magnesium
  • an inorganic acid salt made of hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid or sulfuric acid
  • the pharmaceutical composition of the present invention is prepared in unit dosage form or multi-dose by formulating using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be prepared by incorporating into a container.
  • the pharmaceutically acceptable carrier is one commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like.
  • the pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components.
  • the content of the additives included in the pharmaceutical composition is not particularly limited and may be appropriately adjusted within the range of content used in conventional formulations.
  • the pharmaceutical composition may be formulated as an aqueous solution, suspension, emulsion, etc. for injection, pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma agents. It may be formulated in the form of one or more external preparations selected from the group consisting of agents.
  • the pharmaceutical composition of the present invention may further contain pharmaceutically acceptable carriers and diluents for formulation.
  • pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose, gelatin, alginates, and polyvinyl fibres.
  • binders such as rolidone, etc., lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone, crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol; don't
  • the pharmaceutically acceptable carrier and diluent may be biologically and physiologically compatible with the subject.
  • diluents include, but are not limited to, saline, aqueous buffers, solvents and/or dispersion media.
  • the pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method.
  • parenterally eg, intravenous, subcutaneous, intraperitoneal or topical application
  • it may be formulated into tablets, troches, lozenges, aqueous suspensions, oily suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs.
  • parenteral administration it may be formulated as an injection solution, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.
  • the dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the patient, age, sex, health condition, dietary constitution specificity, the nature of the preparation, the severity of the disease, the administration time of the composition, the administration method, the administration period or interval, and the excretion rate. , And the range may vary depending on the type of drug, and may be appropriately selected by a person skilled in the art. For example, it may be in the range of about 0.1 to 10,000 mg/kg, but is not limited thereto, and may be divided and administered once or several times a day.
  • the pharmaceutical composition may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method.
  • the pharmaceutically effective amount and effective dose of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time and/or administration route of the pharmaceutical composition, and those skilled in the art can use the purpose Dosages effective for treatment can be easily determined and prescribed. Administration of the pharmaceutical composition of the present invention may be administered once a day, or may be divided into several administrations.
  • the present invention provides a preventive or functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and food additives acceptable food additives.
  • ADHD attention deficit hyperactivity disorder
  • the present invention can be generally used as a commonly used food.
  • the food supplement additives include conventional food additives in the art, for example, flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like, and are exemplified below.
  • the food composition of the present invention can be used as a health functional food.
  • health functional food refers to food manufactured and processed using raw materials or ingredients having useful functionalities for the human body in accordance with the Health Functional Food Act, and "functional” refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.
  • the food composition of the present invention may include conventional food additives, and the suitability as the "food additive" is determined according to the general rules of the food additive code approved by the Ministry of Food and Drug Safety and general test methods, etc., unless otherwise specified. It is judged according to the specifications and standards for the item.
  • Items listed in the "Food Additive Code” include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.
  • chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid
  • natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum
  • mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.
  • the food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like.
  • hard capsules can be prepared by mixing and filling the composition according to the present invention with additives such as excipients in conventional hard capsules
  • soft capsules can be prepared by filling the composition according to the present invention. It can be prepared by mixing with additives such as excipients and filling in a capsule base such as gelatin.
  • the soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.
  • prevention refers to all activities that suppress or delay a disease by administering the composition according to the present invention.
  • treatment refers to all activities that improve or beneficially change the symptoms of a disease by administering the composition according to the present invention.
  • improvement means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to a subject.
  • the present invention provides a biomarker composition for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising GAT-3 (GABA transporter-3) or a gene encoding the same as an active ingredient.
  • ADHD attention deficit hyperactivity disorder
  • the present invention provides a diagnostic kit for attention deficit hyperactivity disorder (ADHD) comprising, as an active ingredient, an agent capable of detecting the expression level of GAT-3 (GABA transporter-3) protein or the expression level of a gene encoding the same. to provide.
  • ADHD attention deficit hyperactivity disorder
  • the agent capable of measuring the expression level of the protein is an antibody, peptide, aptamer or compound that binds specifically to the protein, and the agent capable of measuring the expression level of the gene is a primer or a primer that specifically binds to the gene. is a probe
  • Such antibodies include polyclonal antibodies, monoclonal antibodies, recombinant antibodies and complete forms having two full-length light chains and two full-length heavy chains, as well as functional fragments of antibody molecules, such as Fab, F(ab' ), F(ab')2 and Fv.
  • Antibody production can be easily prepared using techniques well known in the field to which the present invention pertains, and commercially available antibodies can be used.
  • protein levels can be measured by immunoassay or immunostaining.
  • the method may be implemented in the form of a microchip or automated microarray system capable of detecting the biomarker protein or fragment thereof in a sample.
  • the immunoassay or immunostaining method may include radioimmunoassay, radioimmunoprecipitation, immunoprecipitation, ELISA, capture-ELISA, inhibition or competition assay, sandwich assay, flow cytometry, immunofluorescence staining, and immunoaffinity purification.
  • the protein level can be measured by multiple reaction monitoring (MRM), parallel reaction monitoring (PRM), sequential windowed data independent acquisition of the total high-resolution (SWATH), and selected reaction monitoring (SRM). ) or using immuno multiple reaction monitoring (iMRM).
  • MRM is a method of determining exact fragments of a substance, breaking them in a mass spectrometer, selecting specific ions among the once-broken ions, and obtaining the number using a continuously connected detector.
  • the 'gene expression level' can be measured using an antisense oligonucleotide, primer pair, or probe that specifically binds to mRNA of the gene, and the agent for measuring the expression of the mRNA is an antisense oligonucleotide specific to the gene. It is selected from the group consisting of nucleotides, primer pairs, probes, and combinations thereof. That is, detection of a nucleic acid may be performed by an amplification reaction using one or more oligonucleotide primers that hybridize to a nucleic acid molecule encoding a gene or a complement of the nucleic acid molecule, but is not limited thereto.
  • mRNA detection using primers can be performed by amplifying a gene sequence using an amplification method such as PCR and then confirming amplification by a method known in the art, such as RT-PCR, competitive RT-PCR, quantitative It can be measured by RT-PCR, RNase protection assay, Northern blot, or DNA chip, but is not limited thereto.
  • an amplification method such as PCR
  • RT-PCR competitive RT-PCR
  • quantitative It can be measured by RT-PCR, RNase protection assay, Northern blot, or DNA chip, but is not limited thereto.
  • the "probe” refers to a nucleic acid fragment such as RNA or DNA corresponding to a few bases to several hundred bases in length that can specifically bind to mRNA, and is labeled to confirm the presence or absence of a specific mRNA and the amount of expression.
  • the probe may be manufactured in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, an RNA probe, or the like. Selection of an appropriate probe and hybridization conditions can be appropriately selected according to techniques known in the art.
  • the "primer” is a nucleic acid sequence having a short free 3' hydroxyl group, capable of forming base pairs with a complementary template and serving as a starting point for template strand copying.
  • Primers can initiate DNA synthesis in the presence of a reagent for polymerization (ie, DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. PCR conditions and lengths of sense and antisense primers can be appropriately selected according to techniques known in the art.
  • the present invention measures the expression level of GAT-3 (GABA transporter-3) protein or the expression level of the gene encoding it in a biological sample isolated from a subject suspected of having attention deficit hyperactivity disorder (ADHD); And it provides an information providing method for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising the step of comparing the expression level with a biological sample isolated from a normal person.
  • GAT-3 GABA transporter-3
  • ADHD attention deficit hyperactivity disorder
  • heterogeneous (+/-) mice lacking the Git1 gene were prepared.
  • a wild-type mouse and a GIT1 gene-defective knock-out mouse are mated to obtain a GIT1 gene-defective heterotype mouse.
  • genotyping was performed to isolate heterotype mice.
  • SNAP5114 was administered by intraperitoneal (i.p) injection at a concentration of 50 ug/kg for 7 days prior to the experiment. SNAP5114 was dissolved in 10% DMSO and then dissolved in 90% Saline immediately before the experiment.
  • mice were perfused. After performing the first perfusion with PBS (Phosphate-buffered saline), 0.05% glutaraldehyde was added to 4% PFA (Paraformaldehyde) to fix it. Thereafter, after collecting the brain from the skull of the mouse, it was immersed in 4% PFA and stored at 4° C. for half a day. The brains of the fixed mice were placed in a 30% sucrose solution and dried for 2 days. After drying, the brain was placed in an OCT compound and stored in a mold at -80°C.
  • PBS Phosphate-buffered saline
  • PFA Paraformaldehyde
  • the hippocampus and striatum were sectioned at a thickness of 30 ⁇ m using a cryotome.
  • the cut slices were washed three times for 5 minutes with PBS, and mixed with triton-X100 and normal goat serum for 1 hour. S100b and GABA were used as primary antibodies and reacted at 4°C for half a day.
  • the cells were washed three times for 5 minutes with PBS, and then reacted with the fluorescently coupled secondary antibody for 2 hours at room temperature. After washing with PBS three times for 5 minutes, the brain tissue was stained on a slide glass using DAKO mounting solution.
  • FIG. 1 The picture on the left of FIG. 1 is a fluorescence picture taken with a confocal microscope after fixing sections of Git1 wild type and heterotype mouse brain striatum and fluorescent tissue immunostaining using a specific antibody.
  • DAPI is for nuclei and s100b is for astrocytes , GABA, gamma amino butyric acid, represents a major inhibitory neurotransmitter in the central nervous system.
  • the graph on the right of FIG. 1 is a confocal micrograph analyzed in Image J.
  • Electrophysiological measurement was performed by cutting the striatum of the mouse at a thickness of 300 um. After bubbling with a mixed gas of 95% O 2 and 5% CO 2 , aCSF (unit mM: 130 NaCl, 24 NaHCO 3 , 1.25 NaH 2 PO 4 , 3.5 KCl, 1.5 CaCl 2 , 1.5 MgCl 2 , and 10 D(+)-glucose, pH 7.4). Measure solution (containing 135 CsCl, 4 NaCl, 0.5 CaCl 2 , 10 HEPES, 5 EGTA, 2 Mg-ATP, 0.5 Na 2 -GTP, and 10 QX-314 adjusted to pH 7.2 with CsOH) into a pipette. Filled to measure the electrophysiological state.
  • aCSF unit mM: 130 NaCl, 24 NaHCO 3 , 1.25 NaH 2 PO 4 , 3.5 KCl, 1.5 CaCl 2 , 1.5 MgCl 2 , and 10 D(+)
  • the upper left corner of FIG. 2 shows a brain slice including the striatum, which is a brain region where patch clamp recording was performed, and a location of the recording pipette.
  • the upper right graph of FIG. 2 shows traces of tonic GABA current measured in medium spiny neurons of the striatum of wild type, hetero type, and knock-out type.
  • the gray bar indicates the induction of full activation current by treatment with GABA
  • the red bar indicates the treatment time with bicuculine, an antagonist of the GABA-A receptor.
  • the base line current is blocked by bicuculine treatment, and the full current and tonic current are measured as indicated by the sky blue arrows.
  • the bottom of Figure 2 is a graph in which the amplitude and frequency of all recorded traces were averaged and significance was analyzed.
  • the tonic GABA current amplitude (pA) on the far right of the bottom was the largest in the wild type and significantly decreased in the heterotype, knock-out type showed the smallest value.
  • the second graph, full activation current (pA), and the third graph, % of full activation, showed similar trends to tonic GABA current.
  • mice were perfused. After performing the first perfusion with PBS (Phosphate-buffered saline), 0.05% glutaraldehyde was added to 4% PFA (Paraformaldehyde) to fix it. Thereafter, after collecting the brain from the skull of the mouse, it was immersed in 4% PFA and stored at 4° C. for half a day. The brains of the fixed mice were placed in a 30% sucrose solution and dried for 2 days. After drying, the brain was placed in an OCT compound and stored in a mold at -80°C.
  • PBS Phosphate-buffered saline
  • PFA Paraformaldehyde
  • the hippocampus and striatum were sectioned at a thickness of 30 ⁇ m using a cryotome.
  • the cut slices were washed three times for 5 minutes with PBS, and mixed with triton-X100 and normal goat serum for 1 hour.
  • S100b and GAT-3 GABA transporter-3) were used as primary antibodies and reacted at 4°C for half a day.
  • the cells were washed three times for 5 minutes with PBS, and then reacted with the fluorescently coupled secondary antibody for 2 hours at room temperature. After washing with PBS three times for 5 minutes, the brain tissue was stained on a slide glass using DAKO mounting solution.
  • the expression level of GAT-3 which is expressed in a large amount in astrocytes, was found to be greatly increased in Git1 heterotype.
  • the bar graph analyzing the intensity of S100b did not show significance, but the intensity of S100b positive GAT-3, that is, GAT3 in astrocytes, was found to increase significantly in Git1 heterotype.
  • GAT-3 expression was analyzed by photographing one astrocyte at the single cell level using IMARIS, and as a result of graph analysis, it was found that GAT-3 expression in Git1 heterotype was significantly increased compared to wild type.
  • FIG. 4 shows the movement of the mouse in the open field and the movement distance and activity level of the mouse.
  • the right side of FIG. 4 shows a graph of the average value of the moving distance of each object used in the experiment.
  • Git1 heterotype (+/-) mice HE
  • SNAP5114 HE SNAP
  • hyperactivity disorder was improved to the level of WT.

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Abstract

The present invention relates to a pharmaceutical composition for preventing or treating attention deficit/hyperactivity disorder (ADHD), comprising SNAP5114 as an active ingredient. A Git1 gene-deficient hetero (+/-) mouse, confirmed to have increased amount of GABA in the brain striatum that controls hyperactivity, can be used as an animal model of ADHD, and by confirming that hyperactivity is ameliorated by administering SNAP5114 to the Git1 gene-deficient hetero (+/-) mouse, SNAP5114 is provided as a therapeutic agent for ADHD.

Description

주의력결핍 과잉행동장애의 예방 또는 치료용 약학적 조성물Pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder
본 발명은 주의력결핍 과잉행동장애(ADHD)의 예방 또는 치료용 약학적 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD).
주의력결핍 과잉행동장애(ADHD)는 유년기에 처음 나타나는 흔한 정신질환 중 하나이며, 이는 또한 성년기에 및 성년기 전반에 걸쳐 발생할 수도 있다. 9 내지 17세 유아의 대략 4.1％가 ADHD를 앓는다는 보고가 있다. ADHD를 앓는 유아는 어떠한 일에 집중해 있을 수 없고, 조용히 앉아 있을 수 없으며, 충동적으로 행동하고, 활동을 끝마칠 수 없다. 이를 치료하지 않으면, 유아의 상해율은 더욱 높아지고, 이 장애는 친구를 사귀는 유아의 능력 및 학교 및(또는) 공부에서의 유아의 직능에 장기적인 부정적 효과를 준다. 시간이 흐르면서, ADHD를 앓는 유아는 우울증, 자존심 박약 및 기타 감정적 문제를 일으킬 가능성이 증가한다.Attention deficit hyperactivity disorder (ADHD) is one of the most common mental disorders that first appear in childhood, and may also occur in and throughout adulthood. It has been reported that approximately 4.1% of children between the ages of 9 and 17 suffer from ADHD. Toddlers with ADHD are unable to concentrate on tasks, cannot sit still, act impulsively, and cannot complete activities. If left untreated, the rate of injury in young children is higher, and the disorder has long-term negative effects on a young child's ability to make friends and on their performance in school and/or studies. Over time, infants with ADHD are more likely to develop depression, low self-esteem and other emotional problems.
대부분의 경우에, ADHD를 앓는 유아 및 성인은 암페타민, 메틸페니데이트 및 페몰린과 같은 정신자극제로 치료한다. 또한, 노르에피네프린의 재흡수를 선택적으로 차단하는 작용을 하는 데심프라민과 같은 항우울제도 일부의 경우에 효과적이다. 또한, 노르에피네프린 및 세로토닌의 재흡수를 차단하는 아토목세틴과 같은 신규 약물도 상기 장애를 치료하는 데에 효과적일 수 있다. 정신자극제 및 모노아민 재흡수 억제제는 활동 수치 및 주의력을 제어하기는 하지만, ADHD와 연관되어서 동반 또는 수반되는 인지 기능 결핍증을 치료하는 데에는 효과적이지 못하다.In most cases, infants and adults with ADHD are treated with psychostimulants such as amphetamines, methylphenidate, and pemoline. In addition, antidepressants such as desipramine, which act to selectively block the reuptake of norepinephrine, are also effective in some cases. Newer drugs such as atomoxetine that block the reuptake of norepinephrine and serotonin may also be effective in treating this disorder. Psychostimulants and monoamine reuptake inhibitors control activity levels and attention, but are ineffective in treating the comorbid or concomitant cognitive deficits associated with ADHD.
ADHD의 원인으로 유전적 요인, 납 수치와 인스턴트식품의 식품첨가물에 대한 반작용으로 보는 생화학적 요인, 뇌에 전달하는 자극을 적절히 선택하는데 문제가 있다고 보는 견해, 환경적 요인 즉 부모와 자녀와의 관계나 부모의 사회적 지위, 임신 중 산모의 흡연과 알콜 남용 등을 언급하기도 하였으나 아직 원인에 대한 논의는 분분하다. 그러나 사회심리적인 요인보다 신경생물학적 요인이 중요하다고 여겨지고 있으며 이에 따라 이 질환의 약물치료와 생물학적 요인에 대한 연구가 활발히 진행되고 있다. 특히 생물학적 요인으로 단일한 신경계의 발달 이상 보다는 고위 인지기능을 담당하는 여러 뇌 영역의 상호연관성의 이상으로 초래되는 비 균일한 질환군일 가능성이 제시되고 있다. 최근까지의 ADHD 환아를 대상으로 한 구조적, 기능적 뇌영상연구를 보면, 대체로 ADHD의 병태생리가 전두-선조회로(fronto-striatal tract)의 기능장애와 연관되어 있으며 메틸페니데이트(Methylphenidate, 이하 MPH)에 의한 약물 효과는 이 영역에서의 도파민계의 기능변화와 연관되어 있다고 알려져 있다. ADHD 증상은 도파민성 신경 외에도 전두엽의 신경회로를 조절하는 노르아드레날린성 신경의 조절과 연관이 있다. ADHD의 행동이 노르아드레 날린성 신경과 도파민성 신경의 조절간에 불균형으로 초래된다는 연구도 보고된 바 있다. 포괄적으로는 Glutamatergic neurons와 GABA (g-aminobutyric acid)성 신경을 지배하는 카테콜라민류가 시냅스 후부에 미치는 영향력이 감소되는데 따른 것으로 이해된다.Genetic factors as causes of ADHD, biochemical factors viewed as a reaction to lead levels and food additives in instant food, view that there is a problem in properly selecting stimuli transmitted to the brain, environmental factors such as the relationship between parents and children The parents' social status, and the mother's smoking and alcohol abuse during pregnancy were also mentioned, but the causes are still debated. However, neurobiological factors are considered to be more important than psychosocial factors, and accordingly, studies on drug treatment and biological factors for this disease are being actively conducted. In particular, it is proposed that it is a non-uniform group of diseases caused by an abnormality in the interconnectivity of various brain regions responsible for high-level cognitive functions rather than a developmental abnormality of a single nervous system as a biological factor. According to structural and functional brain imaging studies targeting children with ADHD, the pathophysiology of ADHD is generally associated with dysfunction of the fronto-striatal tract, and methylphenidate (MPH) ) is known to be associated with changes in the function of the dopamine system in this area. In addition to dopaminergic neurons, ADHD symptoms are related to the control of noradrenergic neurons that control neural circuits in the frontal lobe. Studies have also reported that the behavior of ADHD is caused by an imbalance between the control of noradrenergic and dopaminergic neurons. Comprehensively, it is understood that the influence of catecholamines, which control glutamatergic neurons and GABA (g-aminobutyric acid) neurons, on the post-synaptic area is reduced.
[선행기술문헌][Prior art literature]
[특허문헌][Patent Literature]
한국공개특허공보 제10-2005-0085538호 (2005. 08. 29. 공고)Korean Patent Publication No. 10-2005-0085538 (Announced on August 29, 2005)
본 발명의 목적은 과잉 행동 장애를 개선할 수 있는 효과를 나타내는 성분을 포함하는 주의력결핍 과잉행동장애(ADHD)의 예방 또는 치료용 약학적 조성물을 제공하는 것이다.An object of the present invention is to provide a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) containing a component exhibiting an effect capable of improving hyperactivity disorder.
또한, 본 발명의 다른 목적은 주의력결핍 과잉행동장애(ADHD)를 진단하기 위한 바이오 마커 조성물을 제공하는 것이다.Another object of the present invention is to provide a biomarker composition for diagnosing attention deficit hyperactivity disorder (ADHD).
본 발명은 SNAP5114 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 예방 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) comprising SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient.
또한, 본 발명은 SNAP5114 및 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 주의력결핍 과잉행동장애(ADHD)의 예방 또는 건강기능 식품 조성물을 제공한다.In addition, the present invention provides a preventive or functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and food additives acceptable food additives.
또한, 본 발명은 GAT-3(GABA transporter-3) 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 진단용 바이오 마커 조성물을 제공한다.In addition, the present invention provides a biomarker composition for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising GAT-3 (GABA transporter-3) or a gene encoding the same as an active ingredient.
또한, 본 발명은 GAT-3(GABA transporter-3) 단백질의 발현 수준 또는 이를 코딩하는 유전자의 발현 수준을 검출할 수 있는 제제를 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 진단용 키트를 제공한다.In addition, the present invention provides a diagnostic kit for attention deficit hyperactivity disorder (ADHD) comprising, as an active ingredient, an agent capable of detecting the expression level of GAT-3 (GABA transporter-3) protein or the expression level of a gene encoding the same. to provide.
또한, 본 발명은 주의력결핍 과잉행동장애(ADHD)로 의심되는 대상체로부터 분리된 생물학적 시료에서 GAT-3(GABA transporter-3) 단백질의 발현 수준 또는 이를 코딩하는 유전자의 발현 수준을 측정하는 단계; 및 상기 발현 수준이 정상인으로부터 분리된 생물학적 시료와 비교하는 단계를 포함하는 하는 주의력결핍 과잉행동장애(ADHD)의 진단을 위한 정보 제공 방법을 제공한다.In addition, the present invention measures the expression level of GAT-3 (GABA transporter-3) protein or the expression level of the gene encoding it in a biological sample isolated from a subject suspected of having attention deficit hyperactivity disorder (ADHD); And it provides an information providing method for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising the step of comparing the expression level with a biological sample isolated from a normal person.
본 발명에 따르면, Git1 유전자 결손 헤테로 (+/-) 마우스에서는 과잉행동(hyperactivity)을 관장하는 뇌의 선조체(Striatum)에서 GABA의 양이 증가하는 것을 확인하여 주의력결핍 과잉행동장애(attention deficit / hyperactivity disorder, ADHD) 동물 모델로 사용될 수 있으며, Git1 유전자 결손 헤테로 (+/-) 마우스에 SNAP5114을 투여함에 따라 과잉행동(hyperactivity)이 개선되는 것을 확인함으로써, SNAP5114가 주의력결핍 과잉행동장애(attention deficit / hyperactivity disorder, ADHD)의 치료제로 제공될 수 있고, GAT-3(GABA transporter-3)가 주의력결핍 과잉행동장애(attention deficit / hyperactivity disorder, ADHD)를 진단할 수 있는 바이오 마커로 제공될 수 있다.According to the present invention, it was confirmed that the amount of GABA increased in the striatum of the brain, which is in charge of hyperactivity, in Git1 gene-defective hetero (+/-) mice, thereby preventing attention deficit/hyperactivity disorder. disorder, ADHD) can be used as an animal model, and by confirming that hyperactivity is improved by administering SNAP5114 to Git1 gene-defective hetero (+/-) mice, SNAP5114 can be used as an attention deficit/hyperactivity disorder (ADHD) It can be provided as a treatment for hyperactivity disorder (ADHD), and GABA transporter-3 (GAT-3) can be provided as a biomarker for diagnosing attention deficit/hyperactivity disorder (ADHD).
도 1은 ADHD(attention deficit / hyperactivity disorder) 동물 모델에서 GABA의 양 변화를 형광면역염색(Fluorescent Immunohistochemistry; fIHC)으로 평가한 결과이다.1 is a result of evaluating changes in the amount of GABA in an ADHD (attention deficit / hyperactivity disorder) animal model by fluorescent immunohistochemistry (fIHC).
도 2는 ADHD(attention deficit / hyperactivity disorder) 동물 모델에서 GABA 양의 변화를 전기생리학적 측정(Patch clamp recording)으로 평가한 결과이다.2 is a result of evaluating the change in the amount of GABA in an ADHD (attention deficit / hyperactivity disorder) animal model by electrophysiological measurement (patch clamp recording).
도 3은 ADHD(attention deficit / hyperactivity disorder) 동물 모델에서 SNAP5114 투여에 따른 과잉행동 변화를 평가한 결과이다.3 is a result of evaluating hyperactivity changes according to SNAP5114 administration in an ADHD (attention deficit / hyperactivity disorder) animal model.
본 명세서에서 사용되는 용어는 본 발명에서의 기능을 고려하면서 가능한 현재 널리 사용되는 일반적인 용어들을 선택하였으나, 이는 당 분야에 종사하는 기술자의 의도 또는 판례, 새로운 기술의 출현 등에 따라 달라질 수 있다. 또한, 특정한 경우는 출원인이 임의로 선정한 용어도 있으며, 이 경우 해당되는 발명의 설명 부분에서 상세히 그 의미를 기재할 것이다. 따라서 본 발명에서 사용되는 용어는 단순한 용어의 명칭이 아닌, 그 용어가 가지는 의미와 본 발명의 전반에 걸친 내용을 토대로 정의되어야 한다.The terms used in this specification have been selected from general terms that are currently widely used as much as possible while considering the functions in the present invention, but these may vary depending on the intention of a person skilled in the art, precedent, or the emergence of new technologies. In addition, in a specific case, there is also a term arbitrarily selected by the applicant, and in this case, the meaning will be described in detail in the description of the invention. Therefore, the term used in the present invention should be defined based on the meaning of the term and the overall content of the present invention, not simply the name of the term.
다르게 정의되지 않는 한, 기술적이거나 과학적인 용어를 포함해서 여기서 사용되는 모든 용어들은 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에 의해 일반적으로 이해되는 것과 동일한 의미를 가지고 있다. 일반적으로 사용되는 사전에 정의되어 있는 것과 같은 용어들은 관련 기술의 문맥상 가지는 의미와 일치하는 의미를 가지는 것으로 해석되어야 하며, 본 출원에서 명백하게 정의하지 않는 한, 이상적이거나 과도하게 형식적인 의미로 해석되지 않는다.Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and unless explicitly defined in this application, it should not be interpreted in an ideal or excessively formal meaning. don't
수치 범위는 상기 범위에 정의된 수치를 포함한다. 본 명세서에 걸쳐 주어진 모든 최대의 수치 제한은 낮은 수치 제한이 명확히 쓰여 있는 것처럼 모든 더 낮은 수치 제한을 포함한다. 본 명세서에 걸쳐 주어진 모든 최소의 수치 제한은 더 높은 수치 제한이 명확히 쓰여 있는 것처럼 모든 더 높은 수치 제한을 포함한다. 본 명세서에 걸쳐 주어진 모든 수치 제한은 더 좁은 수치 제한이 명확히 쓰여 있는 것처럼, 더 넓은 수치 범위 내의 더 좋은 모든 수치 범위를 포함할 것이다.Numerical ranges are inclusive of the values defined therein. Every maximum numerical limitation given throughout this specification includes every lower numerical limitation, as if such lower numerical limitations were expressly written. Every minimum numerical limitation given throughout this specification includes every higher numerical limitation, as if such higher numerical limitations were expressly written. Every numerical limitation given throughout this specification will include every better numerical range within the broader numerical range, as if the narrower numerical limitations were expressly written.
이하, 본 발명을 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명은 SNAP5114 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 예방 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) comprising SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient.
상기 SNAP5114는 하기 화학식 1로 표시되는 화합물이며, 구체적으로 (S)-1-(2-(트리스(4-메톡시페닐)메톡시)에틸)피페리딘-3-카르복실산((S)-1-(2-(tris(4-methoxyphenyl)methoxy)ethyl)piperidine-3-carboxylic acid)이다.The SNAP5114 is a compound represented by Formula 1 below, specifically (S)-1-(2-(tris(4-methoxyphenyl)methoxy)ethyl)piperidine-3-carboxylic acid ((S) -1-(2-(tris(4-methoxyphenyl)methoxy)ethyl)piperidine-3-carboxylic acid).
[화학식 1][Formula 1]
Figure PCTKR2021011136-appb-img-000001
Figure PCTKR2021011136-appb-img-000001
상기 SNAP5114는 ADHD(attention deficit / hyperactivity disorder) 동물 모델에서 과잉행동을 개선시키는 효과가 있다.The SNAP5114 has an effect of improving hyperactivity in an attention deficit / hyperactivity disorder (ADHD) animal model.
상기 약학적으로 허용 가능한 염은 약학적으로 허용 가능한 유리산(free acid)에 의해 형성된 산 부가염을 의미하고, 약학적으로 허용 가능한 염은 의약업계에서 통상적으로 사용되는 염을 의미하며, 예를 들어 칼슘, 포타슘, 소듐 또는 마그네슘 등으로 제조된 무기이온염, 염산, 질산, 인산, 브롬산, 요오드산, 과염소산 또는 황산 등으로 제조된 무기산염; 아세트산, 트라이플루오로아세트산, 시트르산, 말레산, 숙신산, 옥살산, 벤조산, 타르타르산, 푸마르산, 만델산, 프로피온산, 젖산, 글리콜산, 글루콘산, 갈락투론산, 글루탐산, 글루타르산, 글루쿠론산, 아스파르트산, 아스코르브산, 카본산 또는, 바닐릭산 등으로 제조된 유기산염; 메탄설폰산, 에탄설폰산, 벤젠설폰산, p-톨루엔설폰산 또는 나프탈렌설폰산 등으로 제조된 설폰산염; 글리신, 아르기닌, 라이신 등으로 제조된 아미노산염; 또는 트라이메틸아민, 트라이에틸아민, 암모니아, 피리딘, 피콜린 등으로 제조된 아민염 등이 있으나, 열거된 이들 염에 의해 본 발명에서 의미하는 염의 종류가 한정되는 것은 아니다.The pharmaceutically acceptable salt refers to an acid addition salt formed by a pharmaceutically acceptable free acid, and the pharmaceutically acceptable salt refers to a salt commonly used in the pharmaceutical industry, for example For example, an inorganic ion salt made of calcium, potassium, sodium or magnesium, an inorganic acid salt made of hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, iodic acid, perchloric acid or sulfuric acid; Acetic acid, trifluoroacetic acid, citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, lactic acid, glycolic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid organic acid salts made of acid, ascorbic acid, carbonic acid or vanillic acid; sulfonic acid salts prepared from methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or naphthalenesulfonic acid; amino acid salts made of glycine, arginine, lysine, and the like; or amine salts prepared with trimethylamine, triethylamine, ammonia, pyridine, picoline, etc., but the types of salts meant in the present invention are not limited by these listed salts.
본 발명의 약학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다.The pharmaceutical composition of the present invention is prepared in unit dosage form or multi-dose by formulating using a pharmaceutically acceptable carrier according to a method that can be easily performed by those skilled in the art. It can be prepared by incorporating into a container.
상기 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸 히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.The pharmaceutically acceptable carrier is one commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components.
본 발명에 있어서, 상기 약학적 조성물에 포함되는 첨가제의 함량은 특별히 한정되는 것은 아니며 통상의 제제화에 사용되는 함량 범위 내에서 적절하게 조절될 수 있다.In the present invention, the content of the additives included in the pharmaceutical composition is not particularly limited and may be appropriately adjusted within the range of content used in conventional formulations.
상기 약학적 조성물은 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립, 정제, 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 및 카타플라스마제로 이루어진 군으로부터 선택되는 하나 이상의 외용제 형태로 제형화될 수 있다.The pharmaceutical composition may be formulated as an aqueous solution, suspension, emulsion, etc. for injection, pills, capsules, granules, tablets, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, and cataplasma agents. It may be formulated in the form of one or more external preparations selected from the group consisting of agents.
본 발명의 약학적 조성물은 제형화를 위해 추가로 있는 약학적으로 허용가능한 담체 및 희석제를 포함할 수 있다. 상기 약학적으로 허용가능한 담체 및 희석제는 전분, 당, 및 만니톨과 같은 부형제, 칼슘 포스페이트 등과 같은 충전제 및 증량제, 카르복시메틸셀룰로오스, 히드록시프로필셀룰로오스 등과 같은 셀룰로오스 유도체, 젤라틴, 알긴산염, 및 폴리비닐 피롤리돈 등과 같은 결합제, 활석, 스테아린산 칼슘, 수소화 피마자유 및 폴리에틸렌 글리콜과 같은 윤활제, 포비돈, 크로스포비돈과 같은 붕해제, 폴리소르베이트, 세틸알코올, 및 글리세롤 등과 같은 계면활성제를 포함하나, 이에 한정되지 않는다. 상기 약학적으로 허용가능한 담체 및 희석제는 대상체에게 생물학적 및 생리학적으로 친화적인 것일 수 있다. 희석제의 예로는 염수, 수용성 완충액, 용매 및/또는 분산제(dispersion media)를 들 수 있으나, 이에 제한되는 것은 아니다.The pharmaceutical composition of the present invention may further contain pharmaceutically acceptable carriers and diluents for formulation. The pharmaceutically acceptable carriers and diluents include excipients such as starch, sugar and mannitol, fillers and extenders such as calcium phosphate, cellulose derivatives such as carboxymethylcellulose and hydroxypropylcellulose, gelatin, alginates, and polyvinyl fibres. binders such as rolidone, etc., lubricants such as talc, calcium stearate, hydrogenated castor oil and polyethylene glycol, disintegrants such as povidone, crospovidone, surfactants such as polysorbates, cetyl alcohol, and glycerol; don't The pharmaceutically acceptable carrier and diluent may be biologically and physiologically compatible with the subject. Examples of diluents include, but are not limited to, saline, aqueous buffers, solvents and/or dispersion media.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구 투여(예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있다. 경구 투여일 경우, 정제, 트로키제 (troches), 로젠지 (lozenge), 수용성 현탁액, 유성 현탁액, 조제 분말, 과립, 에멀젼, 하드 캡슐, 소프트 캡슐, 시럽 또는 엘릭시르제 등으로 제형화될 수 있다. 비경구 투여일 경우, 주사액, 좌제, 호흡기 흡입용 분말, 스프레이용 에어로졸제, 연고, 도포용 파우더, 오일, 크림 등으로 제형화 될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method. For oral administration, it may be formulated into tablets, troches, lozenges, aqueous suspensions, oily suspensions, powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs. In the case of parenteral administration, it may be formulated as an injection solution, suppository, powder for respiratory inhalation, aerosol for spray, ointment, powder for application, oil, cream, etc.
본 발명의 약학적 조성물의 투여량은 환자의 상태 및 체중, 연령, 성별, 건강상태, 식이 체질 특이성, 제제의 성질, 질병의 정도, 조성물의 투여시간, 투여방법, 투여기간 또는 간격, 배설율, 및 약물 형태에 따라 그 범위가 다양할 수 있으며, 이 분야 통상의 기술자에 의해 적절하게 선택될 수 있다. 예컨대, 약 0.1 내지 10,000 mg/kg의 범위일 수 있으나 이제 제한되지 않으며, 하루 일회 내지 수회에 나누어 투여될 수 있다. The dosage of the pharmaceutical composition of the present invention depends on the condition and weight of the patient, age, sex, health condition, dietary constitution specificity, the nature of the preparation, the severity of the disease, the administration time of the composition, the administration method, the administration period or interval, and the excretion rate. , And the range may vary depending on the type of drug, and may be appropriately selected by a person skilled in the art. For example, it may be in the range of about 0.1 to 10,000 mg/kg, but is not limited thereto, and may be divided and administered once or several times a day.
상기 약학적 조성물은 목적하는 방법에 따라 경구 투여되거나 비경구 투여(예를 들면, 정맥 내, 피하 내, 복강 내 또는 국소에 적용)될 수 있다. 본 발명의 약학적 조성물의 약학적 유효량, 유효 투여량은 약학적 조성물의 제제화 방법, 투여 방식, 투여 시간 및/또는 투여 경로 등에 의해 다양해질 수 있으며, 당해 기술 분야에서 통상의 지식을 가진 자는 목적하는 치료에 효과적인 투여량을 용이하게 결정하고 처방할 수 있다. 본 발명의 약학적 조성물의 투여는 하루에 1회 투여될 수 있고, 수회에 나누어 투여될 수도 있다.The pharmaceutical composition may be administered orally or parenterally (eg, intravenous, subcutaneous, intraperitoneal or topical application) depending on the desired method. The pharmaceutically effective amount and effective dose of the pharmaceutical composition of the present invention may vary depending on the formulation method, administration method, administration time and/or administration route of the pharmaceutical composition, and those skilled in the art can use the purpose Dosages effective for treatment can be easily determined and prescribed. Administration of the pharmaceutical composition of the present invention may be administered once a day, or may be divided into several administrations.
또한, 본 발명은 SNAP5114 및 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 주의력결핍 과잉행동장애(ADHD)의 예방 또는 건강기능 식품 조성물을 제공한다.In addition, the present invention provides a preventive or functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and food additives acceptable food additives.
본 발명은 통상적으로 이용되는 식품으로써 일반적으로 사용될 수 있다.The present invention can be generally used as a commonly used food.
상기 식품보조 첨가제는 당업계에 통상적인 식품첨가제, 예를 들어 향미제, 풍미제, 착색제, 충진제, 안정화제 등을 포함하며 하기에 예시한다.The food supplement additives include conventional food additives in the art, for example, flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like, and are exemplified below.
본 발명의 식품 조성물은 건강기능식품으로서 사용될 수 있다. 상기 "건강기능 식품"이라 함은 건강기능 식품에 관한 법률에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.The food composition of the present invention can be used as a health functional food. The term "health functional food" refers to food manufactured and processed using raw materials or ingredients having useful functionalities for the human body in accordance with the Health Functional Food Act, and "functional" refers to food that is not related to the structure and function of the human body. It refers to intake for the purpose of obtaining useful effects for health purposes such as regulating nutrients or physiological functions.
본 발명의 식품 조성물은 통상의 식품 첨가물을 포함할 수 있으며, 상기 "식품 첨가물"로서의 적합 여부는 다른 규정이 없는 한, 식품의약품안전처에 승인된 식품 첨가물 공전의 총칙 및 일반시험법 등에 따라 해당 품목에 관한 규격 및 기준에 의하여 판정한다.The food composition of the present invention may include conventional food additives, and the suitability as the "food additive" is determined according to the general rules of the food additive code approved by the Ministry of Food and Drug Safety and general test methods, etc., unless otherwise specified. It is judged according to the specifications and standards for the item.
상기 "식품 첨가물 공전"에 수재된 품목으로는 예를 들어, 케톤류, 글리신, 구연산칼륨, 니코틴산, 계피산 등의 화학적 합성물, 감색소, 감초추출물, 결정셀룰로오스, 고량색소, 구아검 등의 천연첨가물, L-글루타민산나트륨 제제, 면류첨가알칼리제, 보존료제제, 타르색소제제 등의 혼합제제류들을 들 수 있다.Items listed in the "Food Additive Code" include, for example, chemical compounds such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid, natural additives such as dark pigment, licorice extract, crystalline cellulose, goyang pigment, guar gum, and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations.
본 발명의 식품 조성물은 정제, 캡슐, 분말, 과립, 액상, 환 등의 형태로 제조 및 가공할 수 있다.The food composition of the present invention can be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like.
예를 들어, 캡슐 형태의 건강기능 식품 중 경질캡슐제는 통상의 경질캡슐에 본 발명에 따른 조성물을 부형제 등의 첨가제와 혼합 및 충진하여 제조할 수 있으며, 연질캡슐제는 본 발명에 따른 조성물의 부형제 등의 첨가제와 혼합하고 젤라틴 등 캡슐기제에 충진하여 제조할 수 있다. 상기 연질캡슐제는 필요에 따라 글리세린 또는 소르비톨 등의 가소제, 착색제, 보존제 등을 함유할 수 있다.For example, among the health functional foods in the form of capsules, hard capsules can be prepared by mixing and filling the composition according to the present invention with additives such as excipients in conventional hard capsules, and soft capsules can be prepared by filling the composition according to the present invention. It can be prepared by mixing with additives such as excipients and filling in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.
상기 부형제, 결합제, 붕해제, 활택제, 교미제, 착향제 등에 대한 용어 정의는 당업계에 공지된 문헌에 기재된 것으로 그 기능 등이 동일 내지 유사한 것들을 포함한다. 상기 식품의 종류에는 특별한 제한이 없으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다.Definitions of terms for the excipients, binders, disintegrants, lubricants, corrigents, flavoring agents, etc. are described in literature known in the art, and include those having the same or similar functions. There is no particular limitation on the type of food, and it includes all health functional foods in the usual sense.
본 발명에서 용어 "예방"이란 본 발명에 따른 조성물의 투여로 질환의 억제 또는 지연시키는 모든 행위를 말한다. 본 발명에서 용어 "치료"는 본 발명에 따른 조성물의 투여로 질환의 증세가 호전되거나 이롭게 변경하는 모든 행위를 말한다. 본 발명에서 "개선"이란 본 발명의 조성물을 개체에 투여하거나 섭취시켜 질환의 나쁜 상태를 좋게 하는 모든 행위를 의미한다.In the present invention, the term "prevention" refers to all activities that suppress or delay a disease by administering the composition according to the present invention. In the present invention, the term "treatment" refers to all activities that improve or beneficially change the symptoms of a disease by administering the composition according to the present invention. In the present invention, "improvement" means any action that improves the bad condition of a disease by administering or ingesting the composition of the present invention to a subject.
또한, 본 발명은 GAT-3(GABA transporter-3) 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 진단용 바이오 마커 조성물을 제공한다.In addition, the present invention provides a biomarker composition for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising GAT-3 (GABA transporter-3) or a gene encoding the same as an active ingredient.
또한, 본 발명은 GAT-3(GABA transporter-3) 단백질의 발현 수준 또는 이를 코딩하는 유전자의 발현 수준을 검출할 수 있는 제제를 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 진단용 키트를 제공한다.In addition, the present invention provides a diagnostic kit for attention deficit hyperactivity disorder (ADHD) comprising, as an active ingredient, an agent capable of detecting the expression level of GAT-3 (GABA transporter-3) protein or the expression level of a gene encoding the same. to provide.
상기 단백질의 발현 수준을 측정할 수 있는 제제는 단백질에 특이적으로 결합하는 항체, 펩타이드, 앱타머 또는 화합물이고, 상기 유전자의 발현 수준을 측정할 수 있는 제제는 유전자에 특이적으로 결합하는 프라이머 또는 프로브이다.The agent capable of measuring the expression level of the protein is an antibody, peptide, aptamer or compound that binds specifically to the protein, and the agent capable of measuring the expression level of the gene is a primer or a primer that specifically binds to the gene. is a probe
상기 항체는 다클론 항체, 단클론 항체, 재조합 항체 및 2개의 전체 길이의 경쇄 및 2개의 전체 길이의 중쇄를 가지는 완전한 형태뿐만 아니라 및 항체 분자의 기능적인 단편, 예를 들어, Fab, F(ab'), F(ab')2및 Fv를 모두 포함한다. 항체 생산은 본 발명이 속하는 분야에 널리 공지된 기술을 이용하여 용이하게 제조할 수 있고, 제조되어 상업적으로 판매되는 항체를 이용할 수 있다.Such antibodies include polyclonal antibodies, monoclonal antibodies, recombinant antibodies and complete forms having two full-length light chains and two full-length heavy chains, as well as functional fragments of antibody molecules, such as Fab, F(ab' ), F(ab')2 and Fv. Antibody production can be easily prepared using techniques well known in the field to which the present invention pertains, and commercially available antibodies can be used.
또한, 단백질의 수준은 면역분석법 또는 면역염색법에 의해 측정될 수 있다. 상기 방법은 시료에서 상기 바이오마커 단백질 또는 이의 단편을 검출할 수 있는 마이크로칩 또는 자동화된 마이크로어레이 시스템의 형태로 실시될 수 있다.In addition, protein levels can be measured by immunoassay or immunostaining. The method may be implemented in the form of a microchip or automated microarray system capable of detecting the biomarker protein or fragment thereof in a sample.
상기 면역분석법 또는 면역염색법은 방사능면역분석, 방사능면역침전, 면역침전, ELISA, 캡처-ELISA, 억제 또는 경쟁 분석, 샌드위치 분석, 유세포 분석, 면역형광염색 및 면역친화성 정제를 포함할 수 있다.The immunoassay or immunostaining method may include radioimmunoassay, radioimmunoprecipitation, immunoprecipitation, ELISA, capture-ELISA, inhibition or competition assay, sandwich assay, flow cytometry, immunofluorescence staining, and immunoaffinity purification.
상기 단백질 수준은 다중 반응 모니터링(multiple reaction monitoring: MRM), 병행 반응 모니터링(parallel reaction monitoring: PRM), sequential windowed data independent acquisition of the total high-resolution(SWATH), 선택 반응 모니터링(selected reaction monitoring: SRM) 또는 면역 다중 반응 모니터링(immuno multiple reaction monitoring: iMRM)을 이용하여 측정될 수도 있다. MRM은 물질의 정확한 단편을 결정하여 이를 질량분석기에서 깬 후, 한 번 깨진 이온 중 특정 이온을 한 번 더 선택하여 연속적으로 연결된 검출기를 이용하여 그 수를 얻는 방법이다.The protein level can be measured by multiple reaction monitoring (MRM), parallel reaction monitoring (PRM), sequential windowed data independent acquisition of the total high-resolution (SWATH), and selected reaction monitoring (SRM). ) or using immuno multiple reaction monitoring (iMRM). MRM is a method of determining exact fragments of a substance, breaking them in a mass spectrometer, selecting specific ions among the once-broken ions, and obtaining the number using a continuously connected detector.
상기 '유전자의 발현 수준'은 유전자의 mRNA에 특이적으로 결합하는 안티센스 올리고뉴클레오티드, 프라이머 쌍 또는 프로브를 이용하여 측정할 수 있으며, 상기 mRNA의 발현 여부를 측정하는 제제는 상기 유전자에 특이적인 안티센스 올리고뉴클레오티드, 프라이머 쌍, 프로브 및 이들의 조합으로 이루어진 군에서 선택된다. 즉, 핵산의 검출은 유전자를 암호화하는 핵산 분자 또는 상기 핵산 분자의 상보물에 하이브리드화되는 하나 이상의 올리고뉴클레오타이드 프라이머를 사용하는 증폭반응에 의해 수행될 수 있으나, 이에 제한되는 것은 아니다. 예컨대, 프라이머를 이용한 mRNA의 검출은 PCR과 같은 증폭 방법을 사용하여 유전자 서열을 증폭한 다음 당 분야에 공지된 방법으로 증폭 여부를 확인함으로써 수행될 수 있으며, RT-PCR, 경쟁적 RT-PCR, 정량적 RT-PCR, RNase 보호 분석법, 노던 블롯, 또는 DNA 칩에 의해 측정될 수 있으나, 이에 제한되는 것은 아니다.The 'gene expression level' can be measured using an antisense oligonucleotide, primer pair, or probe that specifically binds to mRNA of the gene, and the agent for measuring the expression of the mRNA is an antisense oligonucleotide specific to the gene. It is selected from the group consisting of nucleotides, primer pairs, probes, and combinations thereof. That is, detection of a nucleic acid may be performed by an amplification reaction using one or more oligonucleotide primers that hybridize to a nucleic acid molecule encoding a gene or a complement of the nucleic acid molecule, but is not limited thereto. For example, mRNA detection using primers can be performed by amplifying a gene sequence using an amplification method such as PCR and then confirming amplification by a method known in the art, such as RT-PCR, competitive RT-PCR, quantitative It can be measured by RT-PCR, RNase protection assay, Northern blot, or DNA chip, but is not limited thereto.
상기 "프로브"는 mRNA외 특이적으로 결합을 이룰 수 있는 짧게는 수 염기 내지 길게는 수백 염기에 해당하는 RNA 또는 DNA 등의 핵산 단편을 의미하며 라벨링되어 있어서 특정 mRNA의 존재 유무, 발현양을 확인할 수 있다. 프로브는 올리고뉴클레오타이드(oligonucleotide) 프로브, 단쇄 DNA(single strand DNA) 프로브, 이중쇄 DNA(double strand DNA) 프로브, RNA 프로브 등의 형태로 제작될 수 있다. 적절한 프로브의 선택 및 혼성화 조건은 당해 기술 분야에 공지된 기술에 따라 적절히 선택할 수 있다.The "probe" refers to a nucleic acid fragment such as RNA or DNA corresponding to a few bases to several hundred bases in length that can specifically bind to mRNA, and is labeled to confirm the presence or absence of a specific mRNA and the amount of expression. can The probe may be manufactured in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, an RNA probe, or the like. Selection of an appropriate probe and hybridization conditions can be appropriately selected according to techniques known in the art.
상기 "프라이머"는 짧은 자유 3-말단 수산화기(free 3' hydroxyl group)를 가지는 핵산 서열로 상보적인 템플레이트(template)와 염기쌍을 형성할 수 있고 템플레이트 가닥 복사를 위한 시작 지점으로서 작용하는 짧은 핵산 서열을 말한다. 프라이머는 적절한 완충용액 및 온도에서 중합반응을 위한 시약(즉, DNA 폴리머라제 또는 역전사효소) 및 상이한 4 가지의 뉴클레오사이드 트리포스페이트의 존재 하에서 DNA 합성을 개시할 수 있다. PCR 조건, 센스 및 안티센스 프라이머의 길이는 당업계에 공지된 기술에 따라 적절히 선택될 수 있다.The "primer" is a nucleic acid sequence having a short free 3' hydroxyl group, capable of forming base pairs with a complementary template and serving as a starting point for template strand copying. say Primers can initiate DNA synthesis in the presence of a reagent for polymerization (ie, DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates in an appropriate buffer and temperature. PCR conditions and lengths of sense and antisense primers can be appropriately selected according to techniques known in the art.
또한, 본 발명은 주의력결핍 과잉행동장애(ADHD)로 의심되는 대상체로부터 분리된 생물학적 시료에서 GAT-3(GABA transporter-3) 단백질의 발현 수준 또는 이를 코딩하는 유전자의 발현 수준을 측정하는 단계; 및 상기 발현 수준이 정상인으로부터 분리된 생물학적 시료와 비교하는 단계를 포함하는 하는 주의력결핍 과잉행동장애(ADHD)의 진단을 위한 정보 제공 방법을 제공한다.In addition, the present invention measures the expression level of GAT-3 (GABA transporter-3) protein or the expression level of the gene encoding it in a biological sample isolated from a subject suspected of having attention deficit hyperactivity disorder (ADHD); And it provides an information providing method for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising the step of comparing the expression level with a biological sample isolated from a normal person.
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 당업계에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.
실시예 1. ADHD 동물 모델 준비Example 1. ADHD animal model preparation
주의력결핍 과잉행동장애(attention deficit / hyperactivity disorder, ADHD) 동물 모델로 Git1 유전자 결손 헤테로 (+/-) 마우스를 준비하였다. 야생형 마우스와 GIT1 유전자 결손 녹-아웃 타입(KO type)인 마우스를 교배하여 GIT1 유전자 결손 헤테로 타입인 마우스를 얻는다. 얻어진 헤테로 타입 마우스끼리의 mating cage를 만든 후, 유전형질분석(genotyping)을 수행하여 헤테로 타입인 마우스를 분리하였다. As an animal model for attention deficit/hyperactivity disorder (ADHD), heterogeneous (+/-) mice lacking the Git1 gene were prepared. A wild-type mouse and a GIT1 gene-defective knock-out mouse are mated to obtain a GIT1 gene-defective heterotype mouse. After creating a mating cage between the obtained heterotype mice, genotyping was performed to isolate heterotype mice.
SNAP5114는 50 ug/kg의 농도로 실험하기 전 7일 동안 복강 주사(intraperitoneal, i.p)로 투여하였다. SNAP5114는 10% DMSO에 녹인 다음, 실험하기 직전에, 90%에 해당하는 Saline에 녹여 사용하였다. SNAP5114 was administered by intraperitoneal (i.p) injection at a concentration of 50 ug/kg for 7 days prior to the experiment. SNAP5114 was dissolved in 10% DMSO and then dissolved in 90% Saline immediately before the experiment.
Open field test를 수행하여 행동학적 형태를 평가하고, 과잉행동(hyperactivity)을 관장하는 대표적인 뇌 부위인 선조체(Striatum)에서 tonic GABA의 특성을 확인하고자 형광면역염색(Immunohistochemistry)과 전기생리학적 측정(patch clamp recording)을 진행하였다.To evaluate behavioral patterns by performing open field tests, and to confirm the characteristics of tonic GABA in the striatum, a representative brain region in charge of hyperactivity, fluorescence immunostaining (Immunohistochemistry) and electrophysiological measurements (patch clamp recording) was performed.
실시예 2. ADHD 동물 모델에서 ADHD 개선 효과 평가Example 2. Evaluation of ADHD improvement effect in ADHD animal model
1) 형광면역염색 (Fluorescent Immunohistochemistry; fIHC)1) Fluorescent Immunohistochemistry (fIHC)
형광면역염색(Fluorescent Immunohistochemistry; fIHC)을 진행하기 위해서, 마우스에서 관류(perfusion)를 진행하였다. PBS(Phosphate-buffered saline)로 1차 관류를 진행한 다음, 4% PFA(Paraformaldehyde)에 0.05%의 글루타르알데히드(glutaraldehyde)를 추가하여 고정하였다. 이후, 마우스의 두개골(skull)에서 뇌를 수집한 후, 4% PFA에 담가 4℃에서 반나절 동안 보관하였다. 고정된 마우스의 뇌는 30% 수크로오스(sucrose) 용액에 넣어 2일 동안 건조 과정을 거쳤다. 건조가 완료된 뇌는 OCT compound에 넣어 -80℃에서 몰드(mold) 형태로 보관하였다. 몰드를 만든 후, 냉동조직절편기(Cryotome)를 이용하여 30 μm의 두께로 해마(hippocampus)와 선상체(striatum)를 절편하였다. 절단한 슬라이스는 PBS로 5분 동안 3회 세척하고, triton-X100과 normal goat serum을 이용하여 1시간 동안 혼합하였다. 일차 항체로 S100b과 GABA 사용하여 4℃에서 반나절 동안 반응시켰다. 항체 반응한 후, PBS로 5분 동안 3회 세척하고, 이후 형광 결합된 2차 항체로 2시간 동안 상온에서 반응시켰다. PBS로 5분 동안 3회 세척하고, DAKO mounting solution을 이용하여 뇌 조직을 슬라이드 글라스에서 염색하였다.To proceed with Fluorescent Immunohistochemistry (fIHC), mice were perfused. After performing the first perfusion with PBS (Phosphate-buffered saline), 0.05% glutaraldehyde was added to 4% PFA (Paraformaldehyde) to fix it. Thereafter, after collecting the brain from the skull of the mouse, it was immersed in 4% PFA and stored at 4° C. for half a day. The brains of the fixed mice were placed in a 30% sucrose solution and dried for 2 days. After drying, the brain was placed in an OCT compound and stored in a mold at -80°C. After making the mold, the hippocampus and striatum were sectioned at a thickness of 30 μm using a cryotome. The cut slices were washed three times for 5 minutes with PBS, and mixed with triton-X100 and normal goat serum for 1 hour. S100b and GABA were used as primary antibodies and reacted at 4°C for half a day. After the antibody reaction, the cells were washed three times for 5 minutes with PBS, and then reacted with the fluorescently coupled secondary antibody for 2 hours at room temperature. After washing with PBS three times for 5 minutes, the brain tissue was stained on a slide glass using DAKO mounting solution.
도 1에 나타난 바와 같이, Git1 hetero type (+/-) 마우스의 성상교세포 내에 있는 GABA의 양이 증가하는 것을 확인하였다. 도 1의 왼쪽 사진은 Git1 wild type과 hetero type 마우스 뇌 선조체 부위 절편을 고정하고, 특이적 항체를 이용한 형광조직면역염색 후에 공초점 현미경으로 촬영한 형광사진으로, DAPI는 핵을, s100b는 성상교세포를, GABA는 gamma amino butyric acid로 중추신경계 주요 억제성 신경전달물질을 나타낸다. 도 1의 오른쪽 그래프는 공초점 현미경 사진을 Image J에서 분석한 것으로, S100b의 intensity를 분석한 바 그래프에서는 유의성이 나타나지 않았으나, S100b positive GABA, 즉, 성상교세포내 GABA의 intensity의 경우 Git1 hetero type에서 유의하게 증가하는 것으로 나타났다. 전체 GABA intensity 및 성상교세포외의 GABA level도 전반적으로 Git1 hetero type에서 증가하는 것으로 나타났다.As shown in FIG. 1, it was confirmed that the amount of GABA in astrocytes of Git1 heterotype (+/-) mice increased. The picture on the left of FIG. 1 is a fluorescence picture taken with a confocal microscope after fixing sections of Git1 wild type and heterotype mouse brain striatum and fluorescent tissue immunostaining using a specific antibody. DAPI is for nuclei and s100b is for astrocytes , GABA, gamma amino butyric acid, represents a major inhibitory neurotransmitter in the central nervous system. The graph on the right of FIG. 1 is a confocal micrograph analyzed in Image J. Significance was not shown in the bar graph where the intensity of S100b was analyzed, but in the case of S100b positive GABA, that is, the intensity of GABA in astrocytes, in Git1 heterotype was found to increase significantly. Total GABA intensity and extra-astrocytic GABA levels were generally increased in Git1 heterotype.
2) 전기생리학적 측정(patch clamp recording)2) Electrophysiological measurement (patch clamp recording)
전기생리학적 측정(Patch clamp recording)은 마우스의 선조체(Striatum)를 300 um의 두께로 절편하여 진행하였다. O2 95%, CO2 5%의 혼합가스로 버블링(bubbling)을 진행한 후에 aCSF (단위 mM: 130 NaCl, 24 NaHCO3, 1.25 NaH2PO4, 3.5 KCl, 1.5 CaCl2, 1.5 MgCl2, 및 10 D(+)-glucose, pH 7.4)에 배양하였다. 측정액(135 CsCl, 4 NaCl, 0.5 CaCl2, 10 HEPES, 5 EGTA, 2 Mg-ATP, 0.5 Na2-GTP, 및 10 QX-314를 포함함. CsOH로 pH 7.2로 맞춤.)을 피펫에 채워 전기생리학적 상태를 측정하였다.Electrophysiological measurement (Patch clamp recording) was performed by cutting the striatum of the mouse at a thickness of 300 um. After bubbling with a mixed gas of 95% O 2 and 5% CO 2 , aCSF (unit mM: 130 NaCl, 24 NaHCO 3 , 1.25 NaH 2 PO 4 , 3.5 KCl, 1.5 CaCl 2 , 1.5 MgCl 2 , and 10 D(+)-glucose, pH 7.4). Measure solution (containing 135 CsCl, 4 NaCl, 0.5 CaCl 2 , 10 HEPES, 5 EGTA, 2 Mg-ATP, 0.5 Na 2 -GTP, and 10 QX-314 adjusted to pH 7.2 with CsOH) into a pipette. Filled to measure the electrophysiological state.
도 2에 나타난 바와 같이, 선조체(Striatum)에서 측정한 tonic GABA의 양이 wild type (+/+)에 비하여 hetero type (+/-)과 knock-out type (-/-)에서 감소하는 경향을 나타냈다. 도 2의 왼쪽 상단은 patch clamp recording을 수행한 뇌 부위인 선조체를 포함하는 절편 (brain slice)과 recording pipette 위치를 나타낸다. 도 2의 오른쪽 상단 그래프는 wild type, hetero type, knock-out type의 선조체의 medium spiny neuron에서 측정한 tonic GABA current의 trace를 나타낸다. 회색 막대는 GABA를 처리해서 Full activation current를 유도한 것이며, 빨간색 막대는 GABA-A 수용체의 antagonist인 bicuculine을 처리한 시간을 나타낸다. bicuculine 처리에 의해 base line current가 blocking 되는데 하늘색 화살표로 나타낸 것처럼 full current와 tonic current를 측정하였다. 도 2의 하단은 기록된 모든 trace들의 amplitude, frequency를 평균 내고 유의성을 분석한 그래프로, 하단 맨 오른쪽 tonic GABA current amplitude (pA)는 wild type에서 가장 크고 hetero type에서 유의하게 감소하였으며, knock-out type이 가장 작은 값을 나타냈다. 두번째 그래프인 full activation current (pA)와 세번째 그래프인 % of full activation의 경우, tonic GABA current와 유사한 경향을 나타냈다.As shown in Figure 2, the amount of tonic GABA measured in the striatum tended to decrease in heterotype (+/-) and knock-out type (-/-) compared to wild type (+/+) showed up The upper left corner of FIG. 2 shows a brain slice including the striatum, which is a brain region where patch clamp recording was performed, and a location of the recording pipette. The upper right graph of FIG. 2 shows traces of tonic GABA current measured in medium spiny neurons of the striatum of wild type, hetero type, and knock-out type. The gray bar indicates the induction of full activation current by treatment with GABA, and the red bar indicates the treatment time with bicuculine, an antagonist of the GABA-A receptor. The base line current is blocked by bicuculine treatment, and the full current and tonic current are measured as indicated by the sky blue arrows. The bottom of Figure 2 is a graph in which the amplitude and frequency of all recorded traces were averaged and significance was analyzed. The tonic GABA current amplitude (pA) on the far right of the bottom was the largest in the wild type and significantly decreased in the heterotype, knock-out type showed the smallest value. The second graph, full activation current (pA), and the third graph, % of full activation, showed similar trends to tonic GABA current.
실시예 3. ADHD 동물 모델에서 GAT-3 수준 평가Example 3. Assessment of GAT-3 levels in ADHD animal models
형광면역염색(Fluorescent Immunohistochemistry; fIHC)을 진행하기 위해서, 마우스에서 관류(perfusion)를 진행하였다. PBS(Phosphate-buffered saline)로 1차 관류를 진행한 다음, 4% PFA(Paraformaldehyde)에 0.05%의 글루타르알데히드(glutaraldehyde)를 추가하여 고정하였다. 이후, 마우스의 두개골(skull)에서 뇌를 수집한 후, 4% PFA에 담가 4℃에서 반나절 동안 보관하였다. 고정된 마우스의 뇌는 30% 수크로오스(sucrose) 용액에 넣어 2일 동안 건조 과정을 거쳤다. 건조가 완료된 뇌는 OCT compound에 넣어 -80℃에서 몰드(mold) 형태로 보관하였다. 몰드를 만든 후, 냉동조직절편기(Cryotome)를 이용하여 30 μm의 두께로 해마(hippocampus)와 선상체(striatum)를 절편하였다. 절단한 슬라이스는 PBS로 5분 동안 3회 세척하고, triton-X100과 normal goat serum을 이용하여 1시간 동안 혼합하였다. 일차 항체로 S100b과 GAT-3 (GABA transporter-3) 사용하여 4℃에서 반나절 동안 반응시켰다. 항체 반응한 후, PBS로 5분 동안 3회 세척하고, 이후 형광 결합된 2차 항체로 2시간 동안 상온에서 반응시켰다. PBS로 5분 동안 3회 세척하고, DAKO mounting solution을 이용하여 뇌 조직을 슬라이드 글라스에서 염색하였다.To proceed with Fluorescent Immunohistochemistry (fIHC), mice were perfused. After performing the first perfusion with PBS (Phosphate-buffered saline), 0.05% glutaraldehyde was added to 4% PFA (Paraformaldehyde) to fix it. Thereafter, after collecting the brain from the skull of the mouse, it was immersed in 4% PFA and stored at 4° C. for half a day. The brains of the fixed mice were placed in a 30% sucrose solution and dried for 2 days. After drying, the brain was placed in an OCT compound and stored in a mold at -80°C. After making the mold, the hippocampus and striatum were sectioned at a thickness of 30 μm using a cryotome. The cut slices were washed three times for 5 minutes with PBS, and mixed with triton-X100 and normal goat serum for 1 hour. S100b and GAT-3 (GABA transporter-3) were used as primary antibodies and reacted at 4°C for half a day. After the antibody reaction, the cells were washed three times for 5 minutes with PBS, and then reacted with the fluorescently coupled secondary antibody for 2 hours at room temperature. After washing with PBS three times for 5 minutes, the brain tissue was stained on a slide glass using DAKO mounting solution.
도 3에 나타난 바와 같이, 성상교세포에서 발현하는 GAT-3 (GABA transporter-3) 단백질의 양을 확인한 결과 GAT-3 단백질의 양은 Wild type (+/+) 마우스에 비해 Git1 hetero type (+/-)에서 높은 수준으로 발현하는 것으로 나타났다. 상기 결과는 주의력결핍 과잉행동장애(attention deficit / hyperactivity disorder, ADHD)가 발병된 마우스에서는 GABA가 성상교세포 안으로 진입하는 수송체인 GAT-3 (GABA transporter-3)가 과도하게 발현되어 성상교세포 내로 GABA양이 증가하는 것을 의미한다.As shown in FIG. 3, as a result of confirming the amount of GAT-3 (GABA transporter-3) protein expressed in astrocytes, the amount of GAT-3 protein was higher in Git1 heterotype (+/-) than in Wild type (+/+) mice. ) was found to be expressed at high levels. The above results show that in mice with attention deficit/hyperactivity disorder (ADHD), GABA transporter-3 (GABA transporter-3), a transporter for GABA entry into astrocytes, is excessively expressed, leading to an increase in the amount of GABA into astrocytes. means that it increases
도 3의 왼쪽에 나타난 바와 같이, 성상교세포에 다량 발현하는 GAT-3의 expression level이 Git1 hetero type에서 크게 증가하는 것으로 나타났다. 도 3의 오른쪽에 나타난 바와 같이, S100b의 intensity를 분석한 바그래프에서는 유의성이 나타나지 않았으나, S100b positive GAT-3, 즉, 성상교세포내 GAT3의 intensity의 경우 Git1 hetero type에서 유의하게 증가하는 것으로 나타났다. IMARIS를 사용하여 성상교세포 하나를 single cell level로 촬영하여 GAT-3 발현을 분석하고, 그래프로 분석한 결과, 통해 Git1 hetero type에서의 GAT-3 발현이 wild type에 비해 유의하게 증가하는 것으로 나타났다.As shown on the left side of Figure 3, the expression level of GAT-3, which is expressed in a large amount in astrocytes, was found to be greatly increased in Git1 heterotype. As shown on the right side of FIG. 3, the bar graph analyzing the intensity of S100b did not show significance, but the intensity of S100b positive GAT-3, that is, GAT3 in astrocytes, was found to increase significantly in Git1 heterotype. GAT-3 expression was analyzed by photographing one astrocyte at the single cell level using IMARIS, and as a result of graph analysis, it was found that GAT-3 expression in Git1 heterotype was significantly increased compared to wild type.
실시예 4. GAT-3 억제제의 처리에 따른 과잉행동 변화Example 4. Hyperactivity changes according to treatment with GAT-3 inhibitors
ADHD 모델 마우스에서의 성상교세포와 연관된 GABA의 양의 변화가 GABA를 성상교세포의 안으로 들여보내는 수송 단백질과 관련이 있음을 고려하여, GAT-3 (GABA transporter-3)의 억제제인 SNAP5114를 처리하여 행동실험을 진행하였다. 행동학적 개선 여부를 평가하기 위해 Open field test를 수행하였다. Open field test를 진행하기 5일 전 handling을 하여, 마우스가 실험자에게 느끼는 스트레스를 최소화하였다. 마우스를 open field test cage(30 X 30 X 30 cm)의 중앙에 넣은 뒤, 10분 동안 움직임을 측정하였다. 측정된 동영상은 ANYmaze를 통하여 locomotor activity를 분석하였다.Considering that the change in the amount of GABA associated with astrocytes in ADHD model mice is related to the transport protein that imports GABA into astrocytes, SNAP5114, an inhibitor of GAT-3 (GABA transporter-3), was treated to act experiment was conducted. An open field test was performed to evaluate behavioral improvement. Handling was performed 5 days before the open field test to minimize the stress felt by the mouse to the experimenter. After putting the mouse in the center of an open field test cage (30 X 30 X 30 cm), movement was measured for 10 minutes. The measured video was analyzed for locomotor activity through ANYmaze.
도 4의 왼쪽은 open field 에서의 마우스 움직임을 추적한 것으로 마우스가 움직인 거리와 활동정도를 나타낸다. 도 4의 오른쪽은 실험에 사용된 각 개체들의 움직인 거리의 평균값을 그래프로 나타낸 것이다. 도 4에 나타난 바와 같이, Git1 hetero type (+/-) 마우스(HE)가 wild type (+/+) 마우스(WT)에 비하여 더 많은 움직임을 나타내는 과잉 행동 장애(hyperactivity)를 나태내는 것을 확인하였다. Git1 hetero type (+/-) 마우스에 SNAP5114를 처리한 경우(HE SNAP) 과잉행동장애가 WT의 수준으로 개선되는 것으로 나타났다.The left side of FIG. 4 shows the movement of the mouse in the open field and the movement distance and activity level of the mouse. The right side of FIG. 4 shows a graph of the average value of the moving distance of each object used in the experiment. As shown in FIG. 4, it was confirmed that Git1 heterotype (+/-) mice (HE) showed hyperactivity showing more movement than wild-type (+/+) mice (WT). . When Git1 heterotype (+/-) mice were treated with SNAP5114 (HE SNAP), hyperactivity disorder was improved to the level of WT.
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 즉, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다.Having described specific parts of the present invention in detail above, it is clear to those skilled in the art that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims (11)

  1. SNAP5114 또는 이의 약학적으로 허용가능한 염을 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 예방 또는 치료용 약학적 조성물.A pharmaceutical composition for preventing or treating attention deficit hyperactivity disorder (ADHD) comprising SNAP5114 or a pharmaceutically acceptable salt thereof as an active ingredient.
  2. 제1항에 있어서, 상기 SNAP5114는 하기 화학식 1로 표시되는 화합물인 것을 특징으로 하는 약학적 조성물.The pharmaceutical composition according to claim 1, wherein the SNAP5114 is a compound represented by Formula 1 below.
    [화학식 1][Formula 1]
    Figure PCTKR2021011136-appb-img-000002
    Figure PCTKR2021011136-appb-img-000002
  3. 제1항에 있어서, 상기 SNAP5114는 과잉행동을 개선시키는 것을 특징으로 하는 약학적 조성물.The pharmaceutical composition according to claim 1, wherein the SNAP5114 improves hyperactivity.
  4. 제1항에 있어서, 상기 약학적 조성물은 제형화를 위해 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸 히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일로 이루어진 군에서 선택된 어느 하나 이상의 담체를 포함하는 것을 특징으로 하는 약학적 조성물.The method of claim 1, wherein the pharmaceutical composition is formulated with lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl A pharmaceutical composition comprising at least one carrier selected from the group consisting of pyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil enemy composition.
  5. SNAP5114 및 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 주의력결핍 과잉행동장애(ADHD)의 예방 또는 건강기능 식품 조성물.A preventive or health functional food composition for attention deficit hyperactivity disorder (ADHD) containing SNAP5114 and a food additive acceptable in food science.
  6. 제5항에 있어서, 상기 SNAP5114는 하기 화학식 1로 표시되는 화합물인 것을 특징으로 하는 건강기능 식품 조성물.The functional food composition according to claim 5, wherein the SNAP5114 is a compound represented by Formula 1 below.
    [화학식 1][Formula 1]
    Figure PCTKR2021011136-appb-img-000003
    Figure PCTKR2021011136-appb-img-000003
  7. GAT-3(GABA transporter-3) 또는 이를 코딩하는 유전자를 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 진단용 바이오 마커 조성물.A biomarker composition for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising GAT-3 (GABA transporter-3) or a gene encoding the same as an active ingredient.
  8. GAT-3(GABA transporter-3) 단백질의 발현 수준 또는 이를 코딩하는 유전자의 발현 수준을 검출할 수 있는 제제를 유효성분으로 포함하는 주의력결핍 과잉행동장애(ADHD)의 진단용 키트.A kit for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising, as an active ingredient, an agent capable of detecting the expression level of a GABA transporter-3 (GAT-3) protein or the expression level of a gene encoding the same.
  9. 제2항에 있어서, 상기 단백질의 발현 수준을 측정할 수 있는 제제는 단백질에 특이적으로 결합하는 항체, 펩타이드, 앱타머 또는 화합물인 것을 특징으로 하는 주의력결핍 과잉행동장애(ADHD)의 진단용 키트.The diagnostic kit for attention deficit hyperactivity disorder (ADHD) according to claim 2, wherein the agent capable of measuring the expression level of the protein is an antibody, peptide, aptamer or compound that specifically binds to the protein.
  10. 제2항에 있어서, 상기 유전자의 발현 수준을 측정할 수 있는 제제는 유전자에 특이적으로 결합하는 프라이머 또는 프로브인 것을 특징으로 하는 주의력결핍 과잉행동장애(ADHD)의 진단용 키트.The diagnostic kit for attention deficit hyperactivity disorder (ADHD) according to claim 2, wherein the agent capable of measuring the expression level of the gene is a primer or probe that specifically binds to the gene.
  11. 주의력결핍 과잉행동장애(ADHD)로 의심되는 대상체로부터 분리된 생물학적 시료에서 GAT-3(GABA transporter-3) 단백질의 발현 수준 또는 이를 코딩하는 유전자의 발현 수준을 측정하는 단계; 및 Measuring the expression level of GABA transporter-3 (GAT-3) protein or the expression level of a gene encoding the same in a biological sample isolated from a subject suspected of having attention deficit hyperactivity disorder (ADHD); and
    상기 발현 수준이 정상인으로부터 분리된 생물학적 시료와 비교하는 단계를 포함하는 하는 주의력결핍 과잉행동장애(ADHD)의 진단을 위한 정보 제공 방법.Information providing method for diagnosis of attention deficit hyperactivity disorder (ADHD) comprising the step of comparing the expression level with a biological sample isolated from a normal person.
PCT/KR2021/011136 2021-08-20 2021-08-20 Pharmaceutical composition for preventing or treating attention deficit/hyperactivity disorder WO2023022269A1 (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20050085538A (en) 2002-12-11 2005-08-29 파마시아 앤드 업존 캄파니 엘엘씨 Combination for the treatment of adhd
WO2005106038A2 (en) * 2004-04-15 2005-11-10 University Of Florida Research Foundation, Inc. Neural proteins as biomarkers for nervous system injury and other neural disorders
KR20120021694A (en) * 2010-08-13 2012-03-09 한국과학기술원 Attention deficit hyperactivity disorder model animal, method for evaluating prevention and alleviation of attention deficit disease, and composition comprising nonspecific t-type calcium channel blocker for prevention and treatment of attention deficit disease

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20050085538A (en) 2002-12-11 2005-08-29 파마시아 앤드 업존 캄파니 엘엘씨 Combination for the treatment of adhd
WO2005106038A2 (en) * 2004-04-15 2005-11-10 University Of Florida Research Foundation, Inc. Neural proteins as biomarkers for nervous system injury and other neural disorders
KR20120021694A (en) * 2010-08-13 2012-03-09 한국과학기술원 Attention deficit hyperactivity disorder model animal, method for evaluating prevention and alleviation of attention deficit disease, and composition comprising nonspecific t-type calcium channel blocker for prevention and treatment of attention deficit disease

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
KERSANTÉ FLAVIE, ROWLEY SAMUEL C. S., PAVLOV IVAN, GUTIÈRREZ‐MECINAS MARÍA, SEMYANOV ALEXEY, REUL JOHANNES M. H. M., WALKER MATTHE: "A functional role for both γ‐aminobutyric acid (GABA) transporter‐1 and GABA transporter‐3 in the modulation of extracellular GABA and GABAergic tonic conductances in the rat hippocampus", THE JOURNAL OF PHYSIOLOGY, WILEY-BLACKWELL PUBLISHING LTD., GB, vol. 591, no. 10, 1 May 2013 (2013-05-01), GB , pages 2429 - 2441, XP093037070, ISSN: 0022-3751, DOI: 10.1113/jphysiol.2012.246298 *
NAAIJEN J, BRALTEN J, POELMANS G, FARAONE STEPHEN, ASHERSON PHILIP, BANASCHEWSKI TOBIAS, BUITELAAR JAN, FRANKE BARBARA, P EBSTEIN : "Glutamatergic and GABAergic gene sets in attention-deficit/hyperactivity disorder: association to overlapping traits in ADHD and autism", TRANSLATIONAL PSYCHIATRY, vol. 7, no. 1, pages e999 - e999, XP093037074, DOI: 10.1038/tp.2016.273 *
YOSHINORI MASUO; MASAMI ISHIDO; MASATOSHI MORITA; HIROFUMI SAWA; KAZUO NAGASHIMA; ETSUO NIKI: "Behavioural characteristics and gene expression in the hyperactive wiggling (Wig) rat", EUROPEAN JOURNAL OF NEUROSCIENCE., OXFORD UNIVERSITY PRESS., GB, vol. 25, no. 12, 28 June 2007 (2007-06-28), GB , pages 3659 - 3666, XP071866205, ISSN: 0953-816X, DOI: 10.1111/j.1460-9568.2007.05613.x *

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